2-methylenetestosterone derivatives



United States Patent O 3,152,153 Z-P/ETHYLENETESTGSTERQNE DEFJVATFJESDavid Eurof Evans, Virginia Water, urrey, and eter John Paimer, Whitton,Engianzl, assignors to Par-lee, Davis 8: Company, Detroit, Mich, acorporation of Michigan No Drawing. Filed Jan. 15, 1962, Ser. No.166,416 Claims priority, application Great Britain Jan. 13, 1961 6Claims. (Cl. 26i)397.4)

oh no wherein Y represents hydrogen or methyl and Z represents ahydroxymethylene, lower alkanoyloxymethylene, lower alkyl substitutedhydroxymethylene, carbonyl, or ethynyl substituted hydroxymethylene,C(Cz l-DOH, group. The hydroxyl or esterified hydroxyl group whenpresent at position 17 has a beta configuration.

In accordance with the invention Z-rnethylene steroids of the aboveformula wherein Z represents hydroxymethylene, lower alkyl substitutedhydroxymethylene or ethynyl substituted hydroxymethylene can be producedby converting the alkoxyoxalyl group in a compound having the formula(H) Y lower alkyl-O O CC wherein R is hydrogen, lower alkyl or ethynyl,and Y is as defined before; or the corresponding enolic formula, to amethylene group. This conversion can be most conveniently carried out byreacting the aforesaid compound with formaldehyde. Formaldehyde per semay be used or it may be created in situ from a polymeric form offormaldehyde such as para-formaldehyde. Preferably an aqueous solutionof formaldehyde is used. The reaction with the formaldehyde is carriedout in the presence of a basic catalyst such as an alkali metalcarbonate or alkaline earth metal carbonate and in a solvent such as alower boiling alcohol like methanol, ethanol, isopropyl alcohol, orother water miscible solvents or mixtures of such solvents with water. Amixture of methanol and water is the preferred solvent medium. Therelative pro portions of the reactants may be varied widely but for bestresults at least one and preferably from thirty to fifty equivalents offormaldehyde should be employed for each equivalent of alkoxyoxalylsteroid. The temperature of the reaction may be varied from C. to 100 C.The reaction is usually carried out either at room temperature or at thereflux temperature of the reaction mixture.

The alkoxyoxalyl steroids employed in the practice of this invention canbe produced by condensing the parent steroids unsubstituted at position2 with an excess r 3,35Z,l53 Ce Patented Get. 6, l fi l of a dialkyloxalate. reactant.

The compounds of the invention wherein Z is an alkanoyloxyrnethylenegroup can be prepared by acylation of the 17-hydroxyl group of a steroidof the formula where Y is as defined before. This is accomplished byracting the 17-hydroxy steroid compound with a lower alkanoic acid or areactive derivative thereof. Some examples of useful acylating agentsare acetic anhydride, acetyl chloride, propionyl chloride, propionicanhydride, and butyryl bromide. For best results, at least oneequivalent and preferably an excess of the acylating agent is employedfor each equivalent of the hydroxy compound. Typically, the acylatingagent serves as solvent for the reaction although inert solvents such asether, benzene, and the like may also be employed. If desired, acylationof the hydroxy compounds can be carried out in the presence of a basesuch as triethylarnine or pyridine. Conveniently, the acylation iscarried out at a tempera ture in the range of from O120 C. However, thetemperature of the reaction is not particularly critical andtemperatures both above and below those set forth are acceptable. Thepreferred temperature range is O to 46 C.

The compounds of the invention wherein Z is a carbonyl group can beproduced by oxidizing the l7-hydroxyl group of a steroid of formulaDiethyl oxalate is a preferred where Y is as defined before, to a ketonegroup. The oxidation is brought about by the action of oxidizing agentscapable of transforming a secondary alcohol group into a keto group.Particularly useful as an oxidizing agent for this reaction is chromiumtrioxide. The relative proportions of the reactants may be varied widelybut it is preferable to employ a slight excess of the oxidizing agent.When one uses chromium trioxide, any excess oxidizing agent can bedestroyed by the addition of methanol to the reaction mixture after theoxidation is complete. Among the various inert solvents suitable for thereaction, acetone is preferred. Temperatures in the range of from 6 toC. are satisfactory and preferably the reaction is carried out between 0and 56 C The products of the invention have useful pharmacologicalproperties. They are anabolic agents exhibiting low androgenic activity.They are pituitary inhibitors and they are also hypocholesteremic agentswhich cause a fall in blood cholesterol without producing the estrogeniceffects normally encountered in the use of such agents. Preferredcompounds with respect to anabolic activity are Z-methylenetestosterone,2methylene-l7a-methyltes tcsterone, Z-methylene-l9nortestosterone,Z-methylene- 17a methyl l-nortestosterone, and2-methylene-l7eethyl-l9-nortestosterone. Preferred compounds withrespect to pituitary inhibition are 2-methyleneandrost-4-ene-3,17-dione, Z-methylene-1'7a-methyltestosterone,2-methylene-i7a-methyl-l9-nortestosterone, and2-methylene-l7aethynyl-l9-nortestosterone. A preferred compound withespect to hypocholesteremic activity is Z-methylenetestosterone. Thecompounds of the invention are effective upon either oral or parenteraladministration.

The following exarnples illustrate the invention.

Example 1 A stirred solution of 28.9 g. of Z-ethoxyoxalyltestosterone in800 ml. methanol and 280 ml. 36% formalin solution is treated dropwiseover a two hour period with a solution of 28.0 g. potassium carbonate in250 ml. water. During tie period of addition, water (400 ml. water) isadded as required to prevent precipitation of the potassium carbonate.Stirring is continued for ninety minutes. The reaction mixture is thendiluted with water and extracted with methylene chloride. The methylenechloride extract is washed first with saturated sodium chloridesolution, then with water, dried over anhydrous sodium sulfate andfinally evaporated to give Z-methylenetestosterone as a yellow oil. Theoil is dissolved in benzene chrmatographed over 400.0 g. of neutralalumina, activity ii. The Z-methylenetestosterone is eluted withbenzene, the eluate containing the desired Z-methylenetestosterone isevaporated to dryness and the residue purified by severalrecrystallizations from acetone; M.P. 160l62 C.

Example 2 A stirred solution of 5.2 g. of 2-ethxyoxalyl-17ctmethyltestosterone in 150 ml. methanol is treated dropwisewith 52 ml. 36% formalin solution. The mixture is sthred for thirtyminutes and then a solution of 5.2 g. potassium carbonate in 80 ml.water is added dropwise over a sixty minute period. Stirring iscontinued for a further ninety minutes, the reaction mixture dilutedwith Water, and extracted with methylene chloride. The methylenechloride extract is washed neutral with water, dried, and evaporated invacuo below 40 C. to give 2- methylene-lh-methy testosterone as a yellowoil. The oil is dissolved in benzene and chromatographed on 150.0 g. ofalumina (Spence type H alumina which has been deactivated with of itsweight of a aqueous acetic acid solution). TheZ-methylene-l7a-methyltestosterone is eluted with benzene, the eluateevaporated to dryness and the residue recrystallized several times fromacetone; M.P. 177-180" C.

Example 3 A stirred solution of 3.5 g. of2-ethoxyoxalyl-17ctethyltestosterone in 105 ml. methanol and ml. 36%formalin solution is treated at room temperature with a solution of 5.0g. potassium carbonate in 35 ml. water over a period of one hour.Stirring is continued for an additional two hours at room temperature,the reaction pour-ed into water and the product extracted with methylenechloride. The extract is washed neutral with water, dried, andevaporated to dryness leaving a residue of 2-methylene-l7ot-ethyltestosterone. The residue is dissolved in benzeneand chromatographed on 150.0 g. of alumina (Spence type H alumina whichhas been deactivated with 5% of its weight of a 10% acetic acidsolution). The Z-methylene-l7a-ethyltestosterone is eluted with benzene,the eluate evaporated to dryness and the residue recrystallized severaltimes from aqueous acetone; M.P. 147-149" C. and 1S6158 C- The2-ethoxyoxalyl-17a ethyltestosterone used as a starting material can beprepared as follows: A stirred solution of 9.0 g. 17a-ethyltestoster0nein 100 ml. anhydrous benzene is treated with 0.681 g. sodium hydridefollowed by 4.15 g. diethyl oxalate. The reaction mixture is stirred atroom temperature for one hour and then at C. for an additional hour.Stirring is then continued at room temperature for a further two hours.Dry ether (200 ml.) is added and the yellow solid is filtered, washed Asolution of 1.5 g. Z-methylenetestosterone in 20 ml. pyridine is treatedwith 10 ml. propionic anhydride. The reaction mixture is allowed tostand overnight at room temperature, poured on ice, acidified andextracted with ether. The ethereal extract is washed with water,saturated sodium carbonate, and finally washed neutral with Water.Evaporation of the ethereal solution to dryness yields the desiredZ-methylenetestosterone propionate. The Z-methylenetestosteronepropionate is chromatographed on g. of alumina (Spence type H aluminawhich has been deactivated with 5% of its weight of a 10% acetic acidsolution). The Z-methylenetestosterone propionate is eluted withpetroleum ether (4060 C.)/ benzene (1:1), the eluate evaporated todryness and the residue crystallized from acetone; M.P. l53l56 C.

By the substitution of 1.5 g. of 2-methylene-19-nortestosterone in theforegoing procedure, the product obtained isZ-methylene-l9-nortestosterone propionate. The corresponding acetic acidesters are obtained by the substitution of acetic anhydride for thepropionic anhydride.

Example 5 A solution of 1.5 g. Z-methylenetestosterone in 200 ml.freshly distilled acetone is treated dropwise at 15 C. with 1.5 ml. ofJones chromic acid reagent. The resulting reaction mixture is stirredfor five minutes, methanol is added to destroy excess chromic acid andmost of the solvent is removed in vacuo. The residue, upon treatmentwith water, gives 2-methyleneandrost-4-ene-3,17-dione as a white solid.This solid is then chromatographed on 100 g. of alumina (Spence type Halumina which has been deactivated with 5% of its weight of a 10% aceticacid solution). The 2 methyleneandrost-4-ene-3,17 dione is eluted withpetroleum ether (40-60" C.)/benzene (1:1), the eluate evaporated todryness and the residue purified by recrystallization from acetone; M.P.198 C.

Example 6 Twenty ml. of 40% formalin solution is slowly added to astirred solution of 2 g. of 2-ethoxyoxalyl-19-nortestosterone in 100 ml.of methanol in a nitrogen atmosphere. After 15 minutes a solution of 2g. of potassium carbonate in 10 ml. of water is added dropwise over a 30minute period. The reaction mixture is stirred for 90 minutes more,poured into saturated sodium chloride solution and the product isextracted with a mixture of ether and methylene chloride. The extract iswashed with water and evaporated to yield about 1.6 g. of a yellow oil.This product in a minimum quantity of benzene is poured on achromatography column containing 45 g. of alumina (Woelm neutralalumina, activity II). The desired product,2-methylene-19-nortestosterone, is obtained by eluting the column with1: 1 benzene-methylene chloride; M.P. 124-126 C. after crystallizationsfrom ether-petroleum ether.

The 2-ethoxyoxalyl-19-nortestosterone used as a starting material can beprepared as follows. A stirred solution of 5.48 g. of 19-nortestosteronein anhydrous benzene is treated at 20 C. under nitrogen with sodiummethoxide solution (prepared from 0.506 g. of sodium and 10 ml. ofmethanol) and reacted with 2.98 ml. of diethyl oxalate over a period of2 /2 hours. The reaction mixture is poured into 600 ml. of anhydrousether and the insoluble yellow product is collected on a filter anddissolved in water. The aqueous solution is acidified and extracted withether. The ether extract is washed with water and evaporated to yield2-ethoxyoxalyl-l9-nortestosterone.

Example 7 A stirred solution of 297 mg. of2-metl1ylene-19-nortestosterone in 30 ml. of acetone at -15 C. istreated with 0.3 ml. of Jones chromic acid reagent (prepared from 26.72g. of chromium trioxide and 23 ml. of concentrated sulfuric acid,diluted to 100 ml. with water) added over a 1 minute period. Stirring iscontinued for an additional 3 minutes and the reaction mixture is pouredinto water and extracted with ether. The ether extract is Washed withWater until neutral, dried, filtered and evaporated to giveZ-methylene-l9-norandrost-4-ene-3,17 dione; M.P. 134- 137 C. aftercrystallizations from petroleum ether.

Example 8 Ten ml. of 40% formalin solution is added to a stirredsolution of 2 g. of Z-ethoxyoxalyl-l7u-methyl-19-nortestosterone in 100ml. of methanol under nitrogen. After minutes a solution of 2 g. ofpotassium carbonate in 5 ml. of water is added and stirring is continuedfor an additional hour. The reaction mixture is poured into saturatedsodium chloride solution and extracted With ether and methylenechloride. The extracts are washed with water until neutral andevaporated to give about 1.56 g. of a yellow foam. This product in aminimum amount of benzene is poured onto a chromatography columncontaining 45 g. alumina (Woelm neutral alumina, activity 111). Thedesired product, 2-methylene-17a-methyl-19- nortestosterone, is elutedfrom the column with benzene;

M.P. 122123 C. after crystallization from acetone-petroleum ether.

The Z-ethoxyoxalyl-l7a-methyl-19-nortestosterone used as a startingmaterial can be prepared as follows. A stirred solution of 5.2 g. of17zx-m6thYl-19-I1OIiCSiOSi6I'OI16 in 90 ml. of anhydrous benzene istreated under nitrogen for 3 hours with 3.4 ml. of diethyl oxalate andsodium methoxide solution (prepared from 0.46 g. of sodium and 10 ml. ofmethanol). Anhydrous ether is added and the insoluble brown solid iscollected on a filter and dissolved in water. The aqueous solution iswashed with benzene, acidified and the white solid product is extractedwith ether. The ether extract is Washed and evaporated to give a residueof 2-ethoxyalyl-17a-methyl-19-nortestosterone.

Example 9 One hundred twenty ml. of 40% formalin solution is added overa 10 minute period to a stirred solution of 12 g. of 2ethoxyoxalyl-17a-ethyl-19-nortestosterone in 900 ml. of methanol undernitrogen. The mixture is stirred for an additional 20 minutes and then asolution of 12 g. of potassium carbonate in 240 ml. of water is addedover a minute period. Stirring is continued for 90 minutes more afterwhich the mixture is diluted with saturated sodium chloride solution andextracted with methylene chloride-ether. The extract is washed withwater until neutral, dried over magnesium sulfate, filtered andevaporated to give about 9 g. of a yellow oil. This oil ischromatographed on alumina 270 g. of neutral alumina, activity 111). Thecolumn is prepared with 99:1 benzene-methanol and eluted with furtherquantities of this solvent to recover the product. If desired, theproduct can be rechromatographed and eluted with 1:1 petroleumether-benzene. It is 2-methylene-l7a-ethyl-l9-nortestosterone, having anultraviolet absorption maximum at 260 millimicrons in ethanol; infraredabsorption maxima in paraffin oil at 3405, 1661, 1615, and 884reciprocal centimeters; [a] =|18 (1.05% in chloroform).

The 2-ethoxyoxalyl-17u-ethyl-19-nortestosterone used as a startingmaterial can be prepared as follows. A solution of 14.5 g. ofl7a-ethyl-l9-nortestosterone in 300 ml. of tertiary butanol is preparedby stirring and warming the mixture to 65 C. After cooling to 60 C. thestirred solution is treated under nitrogen with 13 ml. of diethyloxalate and a solution of 1.65 g. of sodium in 32 n11. of methanol. Thereaction mixture is stirred for 2 more hours and poured into a largevolume of ether. The insoluble solid product (a sodium enolate) iscollected on a filter, washed with ether and dissolved in water. Theaqueous solution is washed with ether-methylene chloride, acidified withcold 4-norma1 hydrochloric acid and extracted with ether. The etherextract obtained in the last operation is washed with water untilneutral, filtered and evaporated to give2-ethoxyoxalyl-17a-ethyl-19-nortestosterone.

Example 10 Sixty-six ml. of 40% formalin solution is added during 30minutes to a stirred solution of 6.8 g. of Z-ethoxyoxalyl 17cc ethynyl19 nortestosterone in 220 ml. of methanol. A solution of 6.6 g. ofpotassium carbonate in 66 ml. of water is added to this mixture over aperiod of one hour and stirring is continued for an additional hour. Thereaction mixture is poured into water, extracted with methylene chlorideand the extract Washed with water, dried and evaporated to give about5.6 g. of a yellow foam. A chromatography column is prepared from g. ofmumina (Woelm neutral alumina, activity HI) with 1:1 petroleumether-benzene. The crude product in the same solvent mixture is pouredonto this column and the desired Z-methylene-l7a-ethynyl-19-nortestosterone is eluted with benzene; M.P. 123-124 C. after severalcrystallizations from acetone-petroleum ether or acetone-hexane.

The Z-ethoxyoxalyl-Hot-ethynyl l9 nortestosterone used as a startingmaterial can be prepared as follows. A solution is prepared by heating9.8 g. of 17a-ethynyl-19 nortestosterone with 295 ml. of tertiarybutanol and stirring at the boiling point until solution is complete.After cooling to 65 C., 8.9 ml. of diethyl oxalate and then a solutionof 1.04 g. of sodium in 20 ml. of methanol is added. The reactionmixture is allowed to cool to room temperature, stirred for 2 more hoursand poured into ether. The sodium enolate compound which is present isextracted with water and the aqueous extract is washed with ether,acidified and extracted with ether. The ether extract obtained in thelast operation is washed with water, dried, filtered and evaporated toyield 2-ethoxyoxalyl-17a-ethyny1-19-nortestosterone; ultravioletabsorption maxima at 246 and 343 millimicrons in ethanol.

By the foregoing procedure, with the substitution of an equivalentamount of Z-ClhOXYOXfilYl-L-8thYl1Ylt6StOS- terone for the 2ethoxyoxalyl-17a-ethynyl-19-nortestosterone, the product obtained isZ-methylene-Not-ethynyltestosterone; M.P. 184 C. The2-ethoxyoxalyl-17uethynyltestosterone can be prepared by the reaction ofl7a-ethynyltestosterone with diethyl oxalate in the general mannerdescribed.

We claim:

1. A compound of the formula where Y is selected from the classconsisting of hydrogen and methyl and Z is selected from the classconsisting of 13 hydroxymethylene, [3 lower alkanoyloxymethylene, loweralkyl substituted {it-hydroxymethylene, carbonyl, and ethynylsubstituted ,B-hydroxymethylene.

2. 2-methylenetestosterone.

3. 2-methylene-17tx-methyltest0sterone.

4. Z-methylene-19-nortestosterone.

5. 2-methylene-l7a-methyl-19-nortestosterone.

6. 2-methylene-17a-ethynyl-19-nortestosterone.

No references cited.

1. A COMPOUND OF THE FORMULA